Integrated liquid chromatography-multiangle light scattering system to measure absolute masses and stoichiometries of biological assemblies in solution

集成液相色谱-多角度光散射系统,用于测量溶液中生物组件的绝对质量和化学计量

基本信息

  • 批准号:
    RTI-2019-00728
  • 负责人:
  • 金额:
    $ 10.92万
  • 依托单位:
  • 依托单位国家:
    加拿大
  • 项目类别:
    Research Tools and Instruments
  • 财政年份:
    2018
  • 资助国家:
    加拿大
  • 起止时间:
    2018-01-01 至 2019-12-31
  • 项目状态:
    已结题

项目摘要

This application requests a state-of-the-art size exclusion chromatography system integrated with multi-angle light scattering (SEC-MALS). SEC-MALS is capable of measuring absolute aggregate molecular weights (MWs) and radii of gyration (Rg) under physiological native solution conditions with unparalleled accuracy, precision and throughput not attainable by conventional analytical SEC. Reliable MW and Rg values are needed to determine the oligomerization state and binding stoichiometry of complexes formed by a wide range of proteins and polymers at the center of a multitude of research programs at McMaster. These include not only globular proteins, but also membrane proteins, intrinsically disordered proteins, non-fouling polymers as well as amyloidogenic polypeptides, which are notoriously refractory to other methods for measuring MW and Rg. The requested instrument, a Wyatt MALS-RI with GE Healthcare SEC, is one of the most versatile and reliable SEC-MALS configurations for academic work currently on the market and will have a transformative impact on a collaborative community of > 8 different NSERC-funded research groups spread across three different faculties at McMaster university, i.e. Faculty of Science, Faculty of Health Sciences and Faculty of Engineering. Currently, there is no SEC-MALS at McMaster. Given the outstanding performance in terms of accuracy, sensitivity and throughput, SEC-MALS is now the gold standard requested by the best international journals for the accurate determination of oligomeric states and macromolecular complex stoichiometries. Without SEC-MALS it is becoming nearly impossible to publish high-impact papers. Furthermore, SEC-MALS has been adopted as the industry standard for the characterization of higher-order structures formed by therapeutic biologicals and polymers. Hence, SEC-MALS will dramatically enhance our ability to train the next generation of HQP in preparation for jobs not only in the academia, but also in the pharma and biotech-sectors. The absence of SEC-MALS at McMaster is severely limiting dozens of researchers and is jeopardizing their ability to maintain internationally competitive research programs. If funded, the requested instrument will be the first SEC-MALS in the whole McMaster campus and will be installed in a newly-renovated centralized, shared and widely accessible core facility. We anticipate an extremely high demand by all groups working in the structural biology, enzymology and bioengineering fields. For virtually all projects in these fields, it is imperative to reliably establish the stoichiometry of homo- and hetero-oligomers before further progress can be achieved and currently SEC-MALS is the prime technique for this purpose. The impact of the research enabled by the requested SEC-MALS ranges from the understanding of the mechanisms of allosteric regulation to the design of new biomaterials. Given the absence of SEC-MALS at McMaster, SEC-MALS is a clear and urgent need. **
该应用程序要求与多角度光散射(SEC-MALS)集成的最先进的排除色谱系统。 SEC-MAL能够以无与伦比的准确性,精度和吞吐量无法通过常规分析SEC来测量生理溶液条件下的绝对骨料分子量(MWS)和回旋的半径(RG)。需要可靠的MW和RG值来确定由McMaster多种研究计划中心的各种蛋白质和聚合物形成的复合物的寡聚状态和结合化的化学计量。这些不仅包括球状蛋白,还包括膜蛋白,内在无序的蛋白质,非污染聚合物以及淀粉样蛋白生成多肽,众所周知,它们对测量MW和RG的其他方法耐火。 The requested instrument, a Wyatt MALS-RI with GE Healthcare SEC, is one of the most versatile and reliable SEC-MALS configurations for academic work currently on the market and will have a transformative impact on a collaborative community of > 8 different NSERC-funded research groups spread across three different faculties at McMaster university, i.e. Faculty of Science, Faculty of Health Sciences and Faculty of Engineering. 目前,麦克马斯特没有SEC-Mals。 鉴于准确性,灵敏度和吞吐量的出色表现,SEC-MALS现在是最佳国际期刊所要求的黄金标准,以准确确定寡聚状态和大分子复合物石化。没有SEC Mals,发表高影响力的论文几乎不可能成为不可能的。此外,SEC-MALS已被作为由治疗生物学和聚合物形成的高阶结构表征的行业标准。因此,SEC-MALS将极大地增强我们培训下一代HQP的能力,不仅在学术界,而且还在制药和生物技术领域做准备工作。麦克马斯特(McMaster)缺乏SEC-MALS严重限制了数十名研究人员,并且正在危害他们维持国际竞争性研究计划的能力。 如果获得资助,请求的仪器将是整个McMaster校园中的第一批SEC-Mals,并将安装在新近装修的集中式,共享且广泛访问的核心设施中。我们预计所有在结构生物学,酶学和生物工程领域中工作的群体的需求极高。 对于几乎所有这些领域的项目,必须可靠地建立同性恋和异性弱体的化学计量,然后才能实现进一步的进展,而目前SEC-MALS是此目的的主要技术。所请求的SEC-MALS所产生的研究的影响范围从对变构调节机制的理解到新生物材料的设计。鉴于麦克马斯特(McMaster)缺乏SEC-Mals,SEC-Mals是一个明显而迫切的需求。 **

项目成果

期刊论文数量(0)
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会议论文数量(0)
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Melacini, Giuseppe其他文献

Aβ Association Inhibition by Transferrin
  • DOI:
    10.1016/j.bpj.2013.03.065
  • 发表时间:
    2013-07-16
  • 期刊:
  • 影响因子:
    3.4
  • 作者:
    Raditsis, Annie V.;Milojevic, Julijana;Melacini, Giuseppe
  • 通讯作者:
    Melacini, Giuseppe
An NMR based phosphodiesterase assay
  • DOI:
    10.1039/d0cc01673j
  • 发表时间:
    2020-07-25
  • 期刊:
  • 影响因子:
    4.9
  • 作者:
    Akimoto, Madoka;Yu, Tianning;Melacini, Giuseppe
  • 通讯作者:
    Melacini, Giuseppe
Mechanism of Selective Enzyme Inhibition through Uncompetitive Regulation of an Allosteric Agonist
  • DOI:
    10.1021/jacs.8b05044
  • 发表时间:
    2018-08-01
  • 期刊:
  • 影响因子:
    15
  • 作者:
    Boulton, Stephen;Selvaratnam, Rajeevan;Melacini, Giuseppe
  • 通讯作者:
    Melacini, Giuseppe
Signaling through dynamic linkers as revealed by PKA
CHESPA/CHESCA-SPARKY: automated NMR data analysis plugins for SPARKY to map protein allostery
CHESPA/CHESCA-SPARKY:SPARKY 的自动化 NMR 数据分析插件,用于绘制蛋白质变构图
  • DOI:
    10.1093/bioinformatics/btaa781
  • 发表时间:
    2020
  • 期刊:
  • 影响因子:
    5.8
  • 作者:
    Shao, Hongzhao;Boulton, Stephen;Olivieri, Cristina;Mohamed, Hebatallah;Akimoto, Madoka;Subrahmanian, Manu Veliparambil;Veglia, Gianluigi;Markley, John L;Melacini, Giuseppe;Lee, Woonghee
  • 通讯作者:
    Lee, Woonghee

Melacini, Giuseppe的其他文献

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{{ truncateString('Melacini, Giuseppe', 18)}}的其他基金

In Situ NMR Studies of Plasma Protein Dynamics and Interactions
血浆蛋白质动力学和相互作用的原位核磁共振研究
  • 批准号:
    RGPIN-2019-05990
  • 财政年份:
    2022
  • 资助金额:
    $ 10.92万
  • 项目类别:
    Discovery Grants Program - Individual
In Situ NMR Studies of Plasma Protein Dynamics and Interactions
血浆蛋白质动力学和相互作用的原位核磁共振研究
  • 批准号:
    RGPIN-2019-05990
  • 财政年份:
    2021
  • 资助金额:
    $ 10.92万
  • 项目类别:
    Discovery Grants Program - Individual
In Situ NMR Studies of Plasma Protein Dynamics and Interactions
血浆蛋白质动力学和相互作用的原位核磁共振研究
  • 批准号:
    RGPIN-2019-05990
  • 财政年份:
    2020
  • 资助金额:
    $ 10.92万
  • 项目类别:
    Discovery Grants Program - Individual
In Situ NMR Studies of Plasma Protein Dynamics and Interactions
血浆蛋白质动力学和相互作用的原位核磁共振研究
  • 批准号:
    RGPIN-2019-05990
  • 财政年份:
    2019
  • 资助金额:
    $ 10.92万
  • 项目类别:
    Discovery Grants Program - Individual
Solution NMR Studies of Interactions of Ligands With Plasma Proteins
配体与血浆蛋白相互作用的溶液核磁共振研究
  • 批准号:
    RGPIN-2014-04514
  • 财政年份:
    2018
  • 资助金额:
    $ 10.92万
  • 项目类别:
    Discovery Grants Program - Individual
Mapping the Effect of Excess Fatty Acids on Albumin-Drug Interactions by NMR
通过 NMR 绘制过量脂肪酸对白蛋白-药物相互作用的影响
  • 批准号:
    521727-2017
  • 财政年份:
    2017
  • 资助金额:
    $ 10.92万
  • 项目类别:
    Engage Grants Program
Solution NMR Studies of Interactions of Ligands With Plasma Proteins
配体与血浆蛋白相互作用的溶液核磁共振研究
  • 批准号:
    RGPIN-2014-04514
  • 财政年份:
    2017
  • 资助金额:
    $ 10.92万
  • 项目类别:
    Discovery Grants Program - Individual
Solution NMR Studies of Interactions of Ligands With Plasma Proteins
配体与血浆蛋白相互作用的溶液核磁共振研究
  • 批准号:
    RGPIN-2014-04514
  • 财政年份:
    2016
  • 资助金额:
    $ 10.92万
  • 项目类别:
    Discovery Grants Program - Individual
Solution NMR Studies of Interactions of Ligands With Plasma Proteins
配体与血浆蛋白相互作用的溶液核磁共振研究
  • 批准号:
    462172-2014
  • 财政年份:
    2016
  • 资助金额:
    $ 10.92万
  • 项目类别:
    Discovery Grants Program - Accelerator Supplements
Mapping the Effect of Lyophilization on the Bone Morphogenetic Protein by NMR
通过 NMR 绘制冻干对骨形态发生蛋白的影响
  • 批准号:
    506923-2016
  • 财政年份:
    2016
  • 资助金额:
    $ 10.92万
  • 项目类别:
    Engage Grants Program

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